AOAC SPIFAN Review Team Meeting Book-July 22, 2015

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Method Validation Report

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Validation Report for the Determination of Thiamin HCl, Riboflavin, Niacin (Nicotinic Acid and Nicotinamide), Pantothenic acid, Vitamin B6 (Pyridoxine, Pyridoxal and Pyridoxamine) and Biotin in Infant Formula and Adult Nutritionals by LC-MS/MS

Document Title

Table 7: Linearity of Pyridoxal Concentration

Area Response 1

Area Response 2

Area Response 3

(ng/ml)

Standard 1 Standard 2 Standard 3 Standard 4 Standard 5 Standard 6 Standard 7 Standard 8 Standard 9 Standard 10 Standard 11 Standard 12 Standard 13 Standard 14 Standard 15 Standard 16 Y-Intercept Correlation Coefficient Acceptance Criteria Slope

0.20 0.40 0.81 1.21 1.61 2.02 2.42 4.84 9.68

0.0305 0.0502 0.0990 0.1575 0.1923 0.2584 0.2915 0.5937 1.1490 1.8090 2.3010 2.8450 l3.2690 4.0130 4.6240 5.2150 0.11825 0.0067

0.0331 0.0547 0.1031 0.1494 0.1976 0.2521 0.2988 0.5826 1.1840 1.6980 2.3860 2.8950 3.3140 3.9200 4.6200

0.0259 0.0514 0.1080 0.1479 0.2023 0.2513 0.2923 0.5890 1.1950 1.7890 2.3510 2.9310 3.4790 3.9570 4.7880 5.2650 0.1208 0.0037

14.53 19.37 24.21 29.05

33.9

38.74 43.58

5.035

(weighting: 1/x)

0.11726 0.00983

0.99967

0.99971

0.9998

Correlation Coefficient (r) ≥ 0.995

Acceptable Y/N

Y

Y

Y

Remark: performed ≥6 point s standard curve to cover working range of product ranges, Correlation Coefficient (r) ≥ 0.995 . Depending on matrix and levels of pyridoxal in the product, range of curve can be selected to decrease number of standards.

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